Red-yellow striped peppers

ABSTRACT

Certain aspects of the present disclosure relate to chimeric pepper plants that produce pepper fruit having a yellow and red striped color, and to seeds that produce the chimeric pepper plants. Other aspects of the present disclosure relate to methods of generating and selecting chimeric pepper plants that produce pepper fruit having a yellow and red striped color, as well as methods of limiting propagation of off-type pepper plant progeny that produce pepper fruit of a single color.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Continuation-In-Part application of U.S.application Ser. No. 14/121,903, filed Oct. 30, 2014, which claims thebenefit of U.S. Provisional Application Ser. No. 61/899,010, filed Nov.1, 2013, and this application also claims the priority benefit of U.S.Provisional Application Ser. No. 62/326,619, filed Apr. 22, 2016, andU.S. Provisional Application Ser. No. 62/366,927, filed Jul. 26, 2016,each of which are incorporated herein by reference in their entirety.

SUBMISSION OF SEQUENCE LISTING ON ASCII TEXT FILE

The content of the following submission on ASCII text file isincorporated herein by reference in its entirety: a computer readableform (CRF) of the Sequence Listing (file name: 701802011300SEQLIST.txt,date recorded: Apr. 19, 2017, size: 10 KB).

FIELD OF THE INVENTION

The present disclosure relates to the field of plant breeding. Inparticular, the present disclosure relates to new and distinctivechimeric pepper plants (Capsicum annuum) that produce yellow and redstriped pepper fruit.

BACKGROUND

The bell pepper (Capsicum annuum) originated in Mexico and theneighboring areas of Central America. Soon after Columbus's discovery ofthis plant, it was grown worldwide and used as a spice and a medicine.Today, pepper plants can be found growing wild in tropical areas aroundthe world. Many countries grow it as a crop. Many of the hot peppers canbe found in Latin America and China, but the United States prefers bellpeppers. Peppers are used for fresh consumption, and they are processedinto powders, sauces, and salsas. Many of the new cultivars grown todaycan be traced back to the early plants.

The genus Capsicum and species annuum includes most of the peppers grownin the United States. These can be further grouped into two broadcategories: chili peppers which are pungent (hot) and sweet pepperswhich are non-pungent (mild). The United States produces four percent ofthe world's Capsicum peppers (sweet and hot), ranking sixth behindChina, Mexico, Turkey, Spain and Nigeria. Bell peppers are the mostcommon sweet pepper and are found in virtually every retail producedepartment. Grown commercially in most states, the U.S. industry islargely concentrated in California and Florida, which together accountedfor 78% of output in 2000. New Jersey, Georgia, and North Carolina roundout the top five producing states (Economic Research Service, USDA,Vegetables and Melons Outlook/VGS-288/Dec. 14, 2001).

Bell peppers are eaten raw, cooked, immature and mature. Oftennutritional content is altered by the changes in the way they areconsumed. Per capita consumption of bell peppers in 1995 was 6.2 pounds.They are an excellent source of Vitamin C, Vitamin A, and Calcium. Redpeppers have more of these qualities than the immature green peppers.

Peppers grown in temperate regions are herbaceous annuals, but areherbaceous perennials where temperatures do not drop below freezing.Pepper plants' growth habit may be prostrate, compact, or erect, but itis determinate in that after it produces nine to eleven leaves a singlestem terminates in flowers. These plants are grown for the edible fleshyfruit produced by this dichotomous growth. Peppers are non-climactericwhich means they do not produce ethylene. They need to stay on the vineto continue the ripening process. A deep taproot will form if the plantroot system is uninjured during transplanting. The spindle root willdevelop fibrous secondary root systems spreading laterally and downward.On the soil surface the stem will produce adventitious roots, but not aseasily as tomatoes. The leaves of the pepper plant arise singly and aresimple, entire, and asymmetrical. Typical of all Solanaceous plants, theleaves are arranged alternately on the stem. They are shiny and glabrousand vary in shape from broadly ovate to ovate lanceolate. The flowersdevelop singly or in twos or threes continuously as the upper structureof the plant proliferates. The corolla is white and five lobed while theanthers are bluish or yellowish in color. The flowers have an openanther formation and will indefinitely self-pollinate. They are alsopollinated by insects, which increases the chances of cross-pollination.Unlike tomatoes, whose pollen becomes nonviable in high temperatures,the pepper flowers' pollen is not extremely heat sensitive and itremains viable up to 100° Fahrenheit producing fruit throughout theseason.

Pepper is an important and valuable field crop. Thus, there is acontinued need for new and interesting peppers.

BRIEF SUMMARY

In order to meet these needs, the present disclosure is directed toimproved hybrid peppers. In certain aspects, the present disclosurerelates to a pepper, Capsicum annuum, seed comprising a yellow alleleand a red allele at the capsanthin-capsorubin synthase (CCS) locus, andproduces a chimeric pepper plant comprising a first tissue, wherein thefirst tissue is an epidermal peel, septum, sepal, petal, anther, or leaftissue comprising only a yellow allele at the CCS locus, and a secondtissue comprising a red allele and a yellow allele at the CCS locus. Incertain embodiments, the second tissue is one or more of filament,placenta, sepal, petal, ovary, anther, vascular, and stem tissue. Insome embodiments, the seed produces a chimeric pepper plant thatproduces pepper fruit having a yellow and red striped color. In someembodiments, the red allele comprises a sequence having at least 90%identity to SEQ ID NO: 1. In some embodiments, the seed produces achimeric pepper plant that produces chimeric pericarp tissue comprisingone or more regions having a yellow color, comprising only a yellowallele at the CCS locus, and one or more regions having a red color,comprising a red allele at the CCS locus. In certain embodiments, theseed is produced from a cross between a first pepper plant that producespepper fruit having a yellow color and second pepper plant that producespepper fruit having a red color. In one embodiment, the first pepperplant is a pepper plant from the ‘OP.1745’ pepper plant variety, and thesecond pepper plant is a pepper plant that produces fruit having a redcolor. In one embodiment, the first pepper plant is a pepper plant fromthe ‘OP.1745’ pepper plant variety, and the second pepper plant is apepper plant from the ‘OP.2023’ pepper plant variety. In anotherembodiment, the first pepper plant is a pepper plant from the ‘OP.1745’pepper plant variety, and the second pepper plant is a pepper plant fromthe ‘OP.1755’ pepper plant variety. In yet another embodiment, the firstpepper plant is a pepper plant from the ‘OP.1745’ pepper plant variety,and the second pepper plant is a pepper plant from the ‘OP.0900’ pepperplant variety.

Certain aspects of the present disclosure relate to Capsicum annuumchimeric pepper plants and plant parts isolated therefrom produced bygrowing any of the pepper seeds described herein. Chimeric pepper plantsof the present disclosure produce pepper fruit having a yellow and redstriped color. In certain embodiments, the present disclosure relates toCapsicum annuum chimeric pepper plants and plant parts isolatedtherefrom having all the physiological and morphological characteristicsof a Capsicum annuum pepper plant produced by growing the pepper seedsdescribed herein. Pepper plant parts include pepper leaves, ovules,pollen, seeds, pepper fruits, parts of pepper fruits, flowers, cells,and the like. In some embodiments, the present disclosure is directed topepper leaves, ovules, pollen, seeds, pepper fruits, parts of pepperfruits, and/or flowers isolated from the chimeric pepper plants. In oneembodiment, the plant part is a pepper fruit. In another embodiment, thepresent disclosure further relates to protoplasts produced from thechimeric pepper plants. In another embodiment, the present disclosurefurther relates to tissue culture of chimeric pepper plants producedfrom a plant part selected from leaf, anther, pistil, stem, petiole,root, root tip, fruit, seed, flower, cotyledon, hypocotyl, embryo andmeristematic cell.

Certain aspects of the present disclosure relate to a method ofgenerating and selecting a chimeric pepper plant that produces pepperfruit having a yellow and red striped color. In some embodiments, themethod includes the steps of crossing a first pepper plant comprisingtwo red alleles at the CCS locus with a second pepper plant comprisingat least one yellow allele at the CCS locus to create F₁ hybrid progeny,growing the F₁ hybrid progeny to produce a population of F₁ hybridprogeny plants, and selecting from the population of F₁ hybrid progenyplants one or more F₁ hybrid progeny plants comprising one or moretissues with only the yellow allele at the CCS locus. In certainembodiments, the one or more selected F₁ hybrid progeny plants producepepper fruit having yellow and red striped color. In certainembodiments, the first pepper plant produces red pepper fruit and thesecond pepper plant produces yellow pepper fruit. In certainembodiments, the first pepper plant is from an ‘OP.2023’ pepper plantvariety, an ‘OP.1755’ pepper plant variety, or an ‘OP.0900’ pepper plantvariety, and the second pepper plant is from an ‘OP.1745’ pepper plantvariety. In certain embodiments, plant material of plants from thepopulation of F₁ hybrid progeny plants is screened to identify plantscontaining one or more tissues with only the yellow allele at the CCSlocus. In one embodiment, the plant material includes any combination ofleaf material, epidermal peel material, and septum material. In oneembodiment, the screening is accomplished by performing singlenucleotide polymorphism (SNP) analysis of the plant material. In certainembodiments, F₁ hybrid progeny plants containing one or more tissueswith only the yellow allele at the CCS locus are selected andpropagated.

Certain aspects of the present disclosure relate to a method ofvegetatively propagating a chimeric pepper plant that produces pepperfruit having a yellow and red striped color, wherein the method includesthe steps of selecting plant material from a chimeric pepper plant thatproduces pepper fruit having a yellow and red striped color, andvegetatively propagating the selected plant material to asexuallyreproduce progeny pepper plants that also produce pepper fruit having ayellow and red striped color. In one embodiment, the plant material issympodial shoot material. In another embodiment, the plant material ismain branch material. In yet another embodiment, the plant material isfrom a branch that produces pepper fruit having a yellow and red stripedcolor. In certain embodiments, the method increases the likelihood ofproducing progeny pepper plants that produce pepper fruit having yellowand red striped color. In one embodiment, the method produces 85% ormore progeny pepper plants that produce pepper fruit having a yellow andred striped color. In other embodiments, the method reduces thelikelihood of producing off-type progeny pepper plants that producepepper fruit having only a single color. In one embodiment, the methodproduces 15% or fewer off-type progeny pepper plants that produce pepperfruit having only a single color.

Other aspects of the present disclosure relate to a method ofvegetatively propagating a chimeric pepper plant that produces pepperfruit having a yellow and red striped color, wherein the method includesthe steps of screening plant material from a chimeric pepper plant thatproduces pepper fruit having a yellow and red striped color, selectingplant material based upon the results of the screening, and vegetativelypropagating the selected plant material to asexually reproduce progenypepper plants that also produce pepper fruit having a yellow and redstriped color. In some embodiments, the screening includes performingsingle nucleotide polymorphism (SNP) analysis on the plant material. Insome embodiments, the plant material is an anther. In certainembodiments, the plant material is selected based upon the plantmaterial being heterozygous for one or more portions of chromosome six.In one embodiment, the one or more portions of chromosome six is theportion spanning position 0.92 Mbp and 180 Mbp on chromosome six. Inanother embodiment, the one or more portions of chromosome six is theportion spanning position 0.92 Mbp and 22 Mbp on chromosome six. In yetanother embodiment, the one or more portions of chromosome six is theportion containing the CCS locus. In certain embodiments, the plantmaterial is selected based upon being heterozygous at the CCS locus. Incertain embodiments, the method increases the likelihood of producingprogeny pepper plants that produce pepper fruit having yellow and redstriped color. In one embodiment, the method produces 85% or moreprogeny pepper plants that produce pepper fruit having a yellow and redstriped color. In other embodiments, the method reduces the likelihoodof producing off-type progeny pepper plants that produce pepper fruithaving only a single color. In one embodiment, the method produces 15%or fewer off-type progeny pepper plants that produce pepper fruit havingonly a single color.

Other aspects of the present disclosure relate to chimeric pepper plantsproduced by any of the methods described herein.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference bystudy of the following descriptions.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawings will be provided by the office upon request and paymentof the necessary fee.

FIG. 1 shows top and side views of several yellow and red striped fruit(RY1).

FIG. 2 shows slices of a yellow and red striped fruit of a chimericpepper plant (RY1).

FIG. 3 shows the marker profiles for markers located on chromosomesfive, six, and seven of the ‘OP.1745’, ‘OP.2023’, ‘Maduro’, and RY1plants. The marker base pair positions are based on the Zunla referencegenome.

FIG. 4 shows a sequence alignment of the portion of thecapsanthin-capsorubin synthase genomic locus for the red (SEQ ID NO: 4)and yellow (SEQ ID NO: 5) alleles used for the CAM000413 markeranalysis. Underlined sequences represent sites for the forward andreverse primers used for PCR amplification of these sequences, andhighlighted nucleotides indicate SNPs detected between the red andyellow alleles.

FIG. 5 shows a side view of a yellow and red striped fruit of a chimericpepper plant (RY2).

FIG. 6 shows a side view of a yellow and red striped fruit of a chimericpepper plant (RY4).

FIGS. 7A-7D shows several yellow and red striped fruit of a chimericpepper plant (RY5), as well as unripened fruit. FIG. 7A shows aside/bottom view of a yellow and red striped fruit of a chimeric pepperplant (RY5). FIG. 7B shows a side view of two yellow and red stripedfruit of a chimeric pepper plant (RY5), as well as an unripened fruit.FIG. 7C shows a side view of two yellow and red striped fruit of achimeric pepper plant (RY5), as well as two unripened fruit. FIG. 7Dshows a side view of two yellow and red striped fruit of a chimericpepper plant (RY5), as well as two unripened fruit.

DETAILED DESCRIPTION

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possess the traits to meetthe program goals. The selected germplasm is crossed in order torecombine the desired traits and through selection varieties or parentlines are developed. The goal is to combine in a single variety orhybrid an improved combination of desirable traits from the parentalgermplasm. These important traits may include higher yield, fieldperformance, fruit and agronomic quality such as fruit shape and length,resistance to diseases and insects, and tolerance to drought and heat.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant cultivars. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Each breeding program should include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended, etc.).

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for three years at least. The best lines are candidatesfor new commercial cultivars; those still deficient in a few traits areused as parents to produce new populations for further selection. Theseprocesses, which lead to the final step of marketing and distribution,usually take from five to ten years from the time the first cross orselection is made.

One goal of pepper plant breeding is to develop new, unique and superiorpepper cultivars. A breeder can initially select and cross two or moreparental lines, followed by repeated selfing and selection, producingmany new genetic combinations. Moreover, a breeder can generate multipledifferent genetic combinations by crossing, selfing, and mutations. Aplant breeder can then select which germplasms to advance to the nextgeneration. These germplasms may then be grown under differentgeographical, climatic, and soil conditions, and further selections canbe made during, and at the end of, the growing season.

The development of commercial pepper cultivars thus requires thedevelopment of pepper parental lines, the crossing of these lines, andthe evaluation of the crosses. Pedigree breeding and recurrent selectionbreeding methods are used to develop cultivars from breedingpopulations. Breeding programs combine desirable traits from two or morevarieties or various broad-based sources into breeding pools from whichlines are developed by selfing and selection of desired phenotypes. Thenew lines are crossed with other lines and the hybrids from thesecrosses are evaluated to determine which have commercial potential.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents which possess favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁s or by intercrossing two F₁s (sib mating). Selection of the bestindividuals is usually begun in the F₂ population; then, beginning inthe F₃, the best individuals in the best families are selected.Replicated testing of families, or hybrid combinations involvingindividuals of these families, often follows in the F₄ generation toimprove the effectiveness of selection for traits with low heritability.At an advanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding may be used to transfer genes for a simply inherited,highly heritable trait into a desirable homozygous cultivar or line thatis the recurrent parent. The source of the trait to be transferred iscalled the donor parent. The resulting plant is expected to have theattributes of the recurrent parent (e.g., cultivar) and the desirabletrait transferred from the donor parent. After the initial cross,individuals possessing the phenotype of the donor parent are selectedand repeatedly crossed (backcrossed) to the recurrent parent. Theresulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. There are many laboratory-based techniques available forthe analysis, comparison and characterization of plant genotype; amongthese are Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length polymorphisms (AFLPs), Simple Sequence Repeats(SSRs—which are also referred to as Microsatellites), and SingleNucleotide Polymorphisms (SNPs).

Molecular markers, or “markers”, can also be used during the breedingprocess for the selection of qualitative traits. For example, markersclosely linked to alleles or markers containing sequences within theactual alleles of interest can be used to select plants that contain thealleles of interest. The use of markers in the selection process isoften called genetic marker enhanced selection or marker-assistedselection. Methods of performing marker analysis are generally known tothose of skill in the art.

Mutation breeding may also be used introducing new traits into peppervarieties. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation (such as X-rays, Gamma rays, neutrons,Beta radiation, or ultraviolet radiation), chemical mutagens (such asbase analogs like 5-bromo-uracil), antibiotics, alkylating agents (suchas sulfur mustards, nitrogen mustards, epoxides, ethyleneamines,sulfates, sulfonates, sulfones, or lactones), azide, hydroxylamine,nitrous acid or acridines. Once a desired trait is observed throughmutagenesis the trait may then be incorporated into existing germplasmby traditional breeding techniques. Details of mutation breeding can befound in Principles of Cultivar Development by Fehr, MacmillanPublishing Company, 1993.

The production of double haploids can also be used for the developmentof homozygous lines in a breeding program. Double haploids are producedby the doubling of a set of chromosomes from a heterozygous plant toproduce a completely homozygous individual. For example, see Wan, etal., Theor. Appl. Genet., 77:889-892, 1989.

Additional non-limiting examples of breeding methods that may be usedinclude, without limitation, those found in Principles of PlantBreeding, John Wiley and Son, pp. 115-161, 1960; Allard, 1960; Simmonds,1979; Sneep et al., 1979; Fehr, 1987; “Carrots and Related VegetableUmbelliferae”, Rubatzky, V. E., et al., 1999.

Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

As used herein, the term “allele” refers to any of one or morealternative forms of a gene, all of which relate to one trait orcharacteristic. In a diploid cell or organism, the two alleles of agiven gene occupy corresponding loci on a pair of homologouschromosomes.

As used herein, the term “zygosity” refers to the degree of similarityof the alleles for a trait. For diploid organisms, a locus may behomozygous (the two alleles are the same), heterozygous (the two allelesare different), and hemizygous (one allele is missing).

As used herein, the term “backcrossing” refers to a process in which abreeder repeatedly crosses hybrid progeny back to one of the parents,for example, a first generation hybrid F₁ with one of the parentalgenotype of the F₁ hybrid.

As used herein, the term “covered cultivation” refers to any type ofcultivation where the plants are not exposed to direct sunlight. Thecovering includes, but is not limited to, greenhouses, glasshouses,net-houses, plastic houses, and tunnels.

As used herein, the term “essentially all the physiological andmorphological characteristics” refers to a plant having thephysiological and morphological characteristics of the recurrent parent.

As used herein, the term “pepper fruit” refers to a fruit produced by aCapsicum annuum plant, commonly referred to as a bell pepper. The colorof a pepper fruit can be green, red, yellow, orange and, more rarely,white, black, and brown, depending on when they are harvested and thespecific cultivar. Green peppers are unripe bell peppers, while theothers are all ripe, with the color variation based on cultivarselection.

As used herein, the term “propagate” refers to reproducing a plant bymeans including, but not limited to, seeds, cuttings, divisions, tissueculture, embryo culture or other in vitro method.

As used herein, the term “regeneration” refers to the development of aplant from tissue culture.

Chimeric Pepper Seeds and Plants

Certain aspects of the present disclosure relate to chimeric pepperplants that produce pepper fruit having a yellow and red striped,patched, and/or variegated color, and to seeds that produce the chimericpepper plants described herein.

Seeds

Certain aspects of the present disclosure relate to one or more seedsthat produce chimeric pepper plants described herein. In someembodiments, the seed comprises a yellow allele and a red allele at thecapsanthin-capsorubin synthase (CCS) locus.

In some embodiments, the seed produces a chimeric pepper plantcomprising a first tissue comprising only a yellow allele at the CCSlocus and a second tissue comprising a red allele and a yellow allele atthe CCS locus. In some embodiments, the first tissue is one or more of eepidermal peel, septum, sepal, petal, anther, or leaf tissue. In someembodiments, the second tissue is one or more of filament, placenta,sepal, petal, anther, ovary, vascular, and stem tissue. In someembodiments, the seed produces a chimeric pepper plant that produceschimeric pericarp having one or more regions having a yellow color andcomprising only a yellow allele at the CCS locus. In some embodiments,the seed produces a chimeric pepper plant that produces chimericpericarp having one or more regions having a red color and comprising ared allele at the CCS locus. In some embodiments, the seed produces achimeric pepper plant that produces chimeric pericarp having one or moreregions having a yellow color and comprising only a yellow allele at theCCS locus and one or more regions having a red color and comprising ared allele at the CCS locus. In some embodiments, the seed produces achimeric pepper plant that produces pepper fruit having a yellow and redstriped, patched, and/or variegated color.

In some embodiments, the seed is produced from a cross between a firstpepper plant and a second pepper plant. In some embodiments, the firstpepper plant is a pepper plant comprising a yellow allele at the CCSlocus. In some embodiments, the first pepper plant is a pepper plantthat produces pepper fruit having a yellow color. In some embodiments,the second pepper plant is a pepper plant comprising a red allele at theCCS locus. In some embodiments, the second pepper plant is a pepperplant that produces pepper fruit having a red color. In someembodiments, the first pepper plant is an ‘OP.1745’ pepper plantvariety. In some embodiments, the second pepper plant is an ‘OP.2023’pepper plant variety, an ‘OP.1755’ pepper plant variety, or an ‘OP.0900’pepper plant variety.

In some embodiments, the seed is produced from a cross between an‘OP.1745’ pepper plant variety and a second pepper plant variety. Insome embodiments, the second pepper plant is a pepper plant comprising ared allele at the CCS locus. In some embodiments, the second pepperplant variety is a pepper plant that produces pepper fruit having a redcolor. In some embodiments, the second pepper plant variety is an‘OP.2023’ pepper plant variety, an ‘OP.1755’ pepper plant variety, or an‘OP.0900’ pepper plant variety.

In some embodiments, the seed is produced from a cross between an‘OP.1745’ pepper plant variety and an ‘OP.2023’ pepper plant variety. Insome embodiments, the seed is produced from a cross between an ‘OP.1745’pepper plant variety and an ‘OP.1755’ pepper plant variety. In someembodiments, the seed is produced from a cross between an ‘OP.1745’pepper plant variety and an ‘OP.0900’ pepper plant variety.

Chimeric Pepper Plants

Certain aspects of the present disclosure relate to chimeric pepperplants that produce pepper fruit having a yellow and red striped,patched, and/or variegated color. In some embodiments, the chimericpepper plants are produced from the pepper seeds described herein.

In some embodiments, the chimeric pepper plant comprises a first tissuecomprising only a yellow allele at the CCS locus and a second tissuecomprising a red allele and a yellow allele at the CCS locus. In someembodiments, the first tissue comprising only a yellow allele at the CCSlocus is one or more of epidermal peel, septum, sepal, petal, anther,and leaf tissue. In some embodiments, the first tissue is epidermal peeltissue. In some embodiments, the first tissue is septum tissue. In someembodiments, the first tissue is sepal tissue. In some embodiments, thefirst tissue is petal tissue. In some embodiments, the first tissue isanther tissue. In some embodiments, the first tissue is leaf tissue. Insome embodiments, the second tissue comprising a red allele and a yellowallele at the CCS locus is one or more of filament, placenta, sepal,petal, ovary, anther, vascular, and stem tissue. In some embodiments,the second tissue is filament tissue. In some embodiments, the secondtissue is placenta tissue. In some embodiments, the second tissue issepal tissue. In some embodiments, the second tissue is petal tissue. Insome embodiments, the second tissue is ovary tissue. In someembodiments, the second tissue is anther tissue. In some embodiments,the second tissue is vascular tissue. In some embodiments, the secondtissue is stem tissue.

In some embodiment, the chimeric pepper plant produces chimeric pericarptissue. In some embodiments, the chimeric pericarp tissue comprises oneor more (e.g., at least one, at least two, at least three, at leastfour, at least five or more) regions having a yellow color. In someembodiments, the chimeric pericarp tissue comprises one or more (e.g.,at least one, at least two, at least three, at least four, at least fiveor more) regions comprising only a yellow allele at the CCS locus. Insome embodiments, the chimeric pericarp tissue comprises one or more(e.g., at least one, at least two, at least three, at least four, atleast five or more) regions having a yellow color and comprising only ayellow allele at the CCS locus. In some embodiments, the chimericpericarp tissue comprises one or more (e.g., at least one, at least two,at least three, at least four, at least five or more) regions having ared color. In some embodiments, the chimeric pericarp tissue comprisesone or more (e.g., at least one, at least two, at least three, at leastfour, at least five or more) regions comprising a red allele at the CCSlocus. In some embodiments, the chimeric pericarp tissue comprises oneor more (e.g., at least one, at least two, at least three, at leastfour, at least five or more) regions having a red color and comprising ared allele at the CCS locus. In some embodiments, the chimeric pericarptissue comprises one or more (e.g., at least one, at least two, at leastthree, at least four, at least five or more) regions having a yellowcolor and one or more (e.g., at least one, at least two, at least three,at least four, at least five or more) regions having a red color. Insome embodiments, the chimeric pericarp tissue comprises one or more(e.g., at least one, at least two, at least three, at least four, atleast five or more) regions comprising only a yellow allele at the CCSlocus and one or more (e.g., at least one, at least two, at least three,at least four, at least five or more) regions comprising a red allele atthe CCS locus. In some embodiments, the chimeric pericarp tissuecomprises one or more regions having a yellow color and comprising onlya yellow allele at the CCS locus and one or more regions having a redcolor and comprising a red allele at the CCS locus. In some embodiments,the chimeric pericarp tissue appears striped, with alternating red andyellow colors. In some embodiments, the chimeric pericarp tissue appearspatched, with alternating red and yellow colors. In some embodiments,the chimeric pericarp tissue appears variegated, with alternating redand yellow colors.

In some embodiments, a chimeric pepper plant of the present disclosureproduces pepper fruit having a yellow and red striped, patched, orvariegated color. In some embodiments, a chimeric pepper plant of thepresent disclosure produces pepper fruit having a yellow and red stripedcolor. In some embodiments, a chimeric pepper plant of the presentdisclosure produces pepper fruit having a yellow and red patched color.In some embodiments, a chimeric pepper plant of the present disclosureproduces pepper fruit having a yellow and red variegated color. In someembodiments, the chimeric pepper plant produces pepper fruit thatappears striped, with alternating yellow and red colors. In someembodiments, the chimeric pepper plant produces pepper fruit thatappears patched, with alternating yellow and red colors. In someembodiments, the chimeric pepper plant produces pepper fruit thatappears variegated, with alternating yellow and red colors.

In some embodiments, the present disclosure relates to one or more plantparts from any of the chimeric pepper plants described herein. In someembodiments, the plant part is a leaf, a seed, a fruit, a cell, or anyportion thereof. In some embodiments, the plant part is a fruit. In someembodiments, the fruit is a pepper fruit having yellow and red striped,patched, and/or variegated colors.

In some embodiments, the present disclosure relates to one or morepollen grains or one or more ovules from any of the chimeric pepperplants described herein. In some embodiments, the present disclosurerelates to a pollen grain from any of the chimeric pepper plantsdescribed herein. In some embodiments, the present disclosure relates toan ovule from any of the chimeric pepper plants described herein.

In some embodiments, the present disclosure relates to one or moreprotoplasts produced from any of the chimeric pepper plants describedherein.

In some embodiments, the present disclosure relates to tissue cultureproduced from protoplasts or cells from any of the chimeric plant pepperplants described herein. In some embodiments, the protoplasts and/orcells are produced from one or more plant parts from any of the chimericplant pepper plants described herein. In some embodiments, the plantpart is one or more of leaf, anther, pistil, stem, petiole, root, roottip, fruit, seed, flower, cotyledon, hypocotyl, embryo, and meristematiccells.

Without wishing to be bound by theory, the chimeric pepper plants of thepresent disclosure may be produced from seed comprising both a yellowallele and a red allele from the parental pepper plants, but some of thetissues in the chimeric pepper plant have lost the parental red allele,while other tissues of the chimeric pepper plant have retained theparental red allele. In some embodiments, the embryo may have formedwith heterogeneous sections of cells, some of which contain only theparental red allele, and some of which contain only the parental yellowallele. These heterogeneous sections of cells may be characterized aspericlinal, mericlinal, or sectorial. In some embodiments, heterogeneoussections of cells may have formed within the meristems of the adultplant, and these sections may be characterized as periclinal,mericlinal, or sectorial. In some embodiments, a mutation in a plastidgene or altered plastid gene expression may cause the chimericphenotype. In some embodiments, altered plastid inheritance patterns maycause the phenotype. In some embodiments, carotenoid synthetic pathwaysor carotenoid accumulation may be affected by plastid alterations,nuclear genome alterations, or developmental perturbations. In someembodiments, the nuclear genome of the hybrid may be incompatible withone parental plastid. In some embodiments, alleles from the maternal andpaternal genomes may interact disharmoniously within the hybrid. In someembodiments, a mutation may be present that inhibits correct mitosis ormeiosis. In some embodiments, a mutation may be present that inhibitscorrect nuclear fusion during fertilization, giving rise to agenetically mosaic embryo. In some embodiments, a transacting factorsuch as a transposon may have removed an allele, a region containing anallele, or a segment of an allele. In some embodiments, a transactingfactor such as a transposon may have integrated into an allele, andcaused the allele to become non-functional. In some embodiments, aportion of the chromosome comprising the red and/or yellow alleles maybe more susceptible to DNA damage than other areas of the pepper genome.In some embodiments, some tissues from the chimeric pepper plant mayhave lost the red allele due to homologous recombination, wherein aportion of the chromosome carrying the yellow allele from one parentalpepper plant replaces the portion of the chromosome carrying the redallele from the other parental pepper plant. In some embodiments, thetissues having lost the red allele due to homologous recombination wouldbe homozygous for at least a portion of the parental chromosome carryingthe yellow allele. In some embodiments, some tissues from the chimericpepper plant may have lost at least a portion of the chromosomescomprising the red allele due to genomic instability. In someembodiments, loss of at least a portion of the chromosome carrying thered allele may lead to tissues that are hemizygous for at least aportion of the parental chromosome carrying the yellow allele. In someembodiments, the genomic instability may be due to genomic instabilityon the chromosome comprising the red or yellow alleles. In someembodiments, the genomic instability may be on a chromosome that doesnot comprise the red or yellow alleles, but induces loss of at least aportion of the chromosome carrying the red allele. In some embodiments,the genomic instability may be due to a transacting factor, such as atransposon, causing instability on the chromosome carrying the red oryellow alleles. In some embodiments, the genomic instability may be dueto invasion by a foreign element, such as a virus.

Red and Yellow CCS Alleles

In some embodiments, a pepper seed, tissue, organ, plant part, and/orplant comprising a red allele at the CCS locus is a pepper seed, tissue,organ, plant part, and/or plant with a genomic locus comprising apolynucleotide of SEQ ID NO: 1. In some embodiments, a pepper seed,tissue, organ, plant part, and/or plant comprising a red allele at theCCS locus is a pepper seed, tissue, organ, plant part, and/or plant witha genomic locus comprising a polynucleotide having at least 90%, atleast 91%, at least 92%, at least 93%, at least 94%, at least 95%, atleast 96%, at least 97%, at least 98%, at least 99%, or 100% identity tothe sequence of SEQ ID NO: 1.

In some embodiments, a pepper seed, tissue, organ, plant part, and/orplant comprising a red allele at the CCS locus is a pepper seed, tissue,organ, plant part, and/or plant which encodes a polynucleotide of SEQ IDNO: 2. In some embodiments, a pepper seed, tissue, organ, plant part,and/or plant comprising a red allele at the CCS locus is a pepper seed,tissue, organ, plant part, and/or plant which encodes a polynucleotidehaving at least at least 90%, at least 91%, at least 92%, at least 93%,at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to the sequence of SEQ ID NO: 2.

In some embodiments, a pepper seed, tissue, organ, plant part, and/orplant comprising a red allele at the CCS locus is a pepper seed, tissue,organ, plant part, and/or plant comprising a polypeptide of SEQ ID NO:3. In some embodiments, a pepper seed, tissue, organ, plant part, and/orplant comprising a red allele at the CCS locus is a pepper seed, tissue,organ, plant part, and/or plant comprising a polypeptide having at least90%, at least 91%, at least 92%, at least 93%, at least 94%, at least95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%identity to the sequence of SEQ ID NO: 3.

In some embodiments, a pepper seed, tissue, organ, plant part, and/orplant comprising a yellow allele at the CCS locus is a pepper seed,tissue, organ, plant part, and/or plant that produces a less functionalor non-functional CCS gene product. In some embodiments, a pepper seed,tissue, organ, plant part, and/or plant that produces a less functionalor non-functional CCS gene product is a pepper seed, tissue, organ,plant part, and/or plant comprising an inactivating mutation in the CCSgene. Examples of inactivating mutations may include, but are notlimited to, point mutations, nonsense mutations, truncation mutations,missense mutations, substitution mutations, frameshift mutations,loss-of-function mutations, deletion mutations, insertion mutations,duplication mutations, amplification mutations, translocation mutations,or inversion mutations that result in a non-functional gene productencoded by the gene. In some embodiments, a pepper seed, tissue, organ,plant part, and/or plant that produces a less functional ornon-functional CCS gene product is a pepper seed, tissue, organ, plantpart, and/or plant comprising epigenetic modification to the CCS genelocus. In some embodiments, epigenetic modification to the CCS locusreduces or inhibits expression of the CCS gene product. In someembodiments, a pepper seed, tissue, organ, plant part, and/or plant thatproduces a less functional or non-functional CCS gene product is apepper seed, tissue, organ, plant part, and/or plant comprising CCS geneproduct that has been inactivated. Examples of inactivated gene productsmay include, but are not limited to, gene products that are misfolded,gene products that are inhibited by post-translational modifications,gene products inhibited by binding to inhibitors, gene products that arechemically inhibited, and gene products that are mislocalized.

Methods of Generating and Selecting Chimeric Pepper Plants

Certain aspects of the present disclosure relate to methods ofgenerating and selecting chimeric pepper plants that produce pepperfruit having a yellow and red striped, patched, and/or variegated color.

In some embodiments, the method of generating and selecting pepperplants that produce pepper fruit having a yellow and red striped,patched, and/or variegated color comprises crossing a first parentalpepper plant and a second parental pepper plant. In some embodiments,the first parental pepper plant is the male parental pepper plant andthe second parental pepper plant is the female parental pepper plant. Insome embodiments, the first parental pepper plant is the female parentalpepper plant and the second parental pepper plant is the male parentalpepper plant. In some embodiments, the first parental pepper plant andthe second pepper plant are plants from inbred pepper plant lines.

In some embodiments, the first parental pepper plant comprises at leastone (e.g., one, two, or more) red alleles (R) at the CCS locus. In someembodiments, the first parental pepper plant comprises two red alleles(RR) at the CCS locus. In some embodiments, the first parental pepperplant comprises an (RR) genotype. In some embodiments, the firstparental pepper plant comprises one red allele (R) and one yellow allele(r) at the CCS locus. In some embodiments, the first parental pepperplant comprises two red alleles (Rr) genotype. In some embodiments, thesecond parental pepper plant comprises at least one (e.g., one, two, ormore) yellow alleles (r) at the CCS locus. In some embodiments, thesecond parental pepper plant comprises two yellow alleles (rr) at theCCS locus. In some embodiments, the second parental pepper plantcomprises an (rr) genotype. In some embodiments, the second parentalpepper plant comprises one yellow allele (r) and one red allele (R) atthe CCS locus. In some embodiments, the second parental pepper plantcomprises an (Rr) genotype.

In some embodiments, the method of generating chimeric pepper plantscomprises crossing a first parental pepper plant comprising an (RR) or(Rr) genotype with a second parental pepper plant comprising an (Rr) or(rr) genotype to produce hybrid seed. In some embodiments, the firstparental pepper plant comprises an (RR) genotype and the second parentalpepper plant comprises an (Rr) genotype. In some embodiments, the firstparental pepper plant comprises an (RR) genotype and the second parentalpepper plant comprises an (rr) genotype. In some embodiments, the firstparental pepper plant comprises an (Rr) genotype and the second parentalpepper plant comprises an (Rr) genotype. In some embodiments, the firstparental pepper plant comprises an (Rr) genotype and the second parentalpepper plant comprises an (rr) genotype.

In some embodiments, the first parental pepper plant is an ‘OP.2023’pepper plant variety, an ‘OP.1755’ pepper plant variety, or an ‘OP.0900’pepper plant variety. In some embodiments, the first parental pepperplant is an ‘OP.2023’ pepper plant variety. In some embodiments, thefirst parental pepper plant is an ‘OP.1755’ pepper plant variety. Insome embodiments, the first parental pepper plant is an ‘OP.0900’ pepperplant variety. In some embodiments, the second parental pepper plant isan ‘OP.1745’ pepper plant variety. In some embodiments, the crossbetween the first parental pepper plant and the second parental pepperplant comprises a cross between an ‘OP.2023’ pepper plant variety and an‘OP.1745’ pepper plant variety. In some embodiments, the cross betweenthe first parental pepper plant and the second parental pepper plantcomprises a cross between an ‘OP.1755’ pepper plant variety and an‘OP.1745’ pepper plant variety. In some embodiments, the cross betweenthe first parental pepper plant and the second parental pepper plantcomprises a cross between an ‘OP.0900’ pepper plant variety and an‘OP.1745’ pepper plant variety.

In some embodiments, a cross between the first parental pepper plant andsecond parental pepper plant generates F₁ hybrid progeny. In someembodiments, the F₁ hybrid progeny is grown to create a population of F₁hybrid progeny plants. In some embodiments, one or more F₁ hybridprogeny plants that produce pepper fruit having a yellow and red stripedcolor are selected from the population of F₁hybrid progeny plants. Insome embodiments, the one or more selected F₁hybrid progeny plantscomprise one or more tissues comprising only a yellow allele at the CCSlocus. In some embodiments, plant material from the population ofF₁hybrid progeny plants is screened to identify F₁ hybrid progeny plantscomprising one or more tissues comprising only a yellow allele at theCCS locus. In some embodiments, the plant material is one or more ofleaf, epidermal peel, and septum material. In some embodiments, theplant material is leaf material. In some embodiments, the plant materialis epidermal peel material. In some embodiments, the plant material isseptum material.

In some embodiments, screening plant material comprises performingmarker analysis on the plant material. In some embodiments, the markeranalysis is single nucleotide polymorphism (SNP) analysis. In someembodiments, the marker analysis indicates the zygosity of one or morechromosomes in the plant material. In some embodiments, the markeranalysis indicates the zygosity of chromosome six in the plant material.Examples of markers on chromosome six may include, but are not limitedto, CONTIG14444:607, CONTIG3495:1013, CONTIG13973:146, CONTIG15080:278,CAM0000501, CONTIG200085:278, CONTIG1349:1399, CONTIG5713:302,CONTIG1355:1037, CONTIG11146:187, CONTIGCAM0002444, CONTIG8615:1817,CONTIG22248:401, CONTIG3916:1756, CONTIG7457:106, CAM0003794,CONTIG3740:1525, CONTIG462:888, CONTIG2594:540, CAM000413, and themarkers described in Lefebvre, V. et al. Plant Mol Biol 1998 36(5)785-789. In some embodiments, the marker is CAM000413. In someembodiments, the marker analysis indicates the zygosity of one or moreportions of chromosome six in the plant material. In some embodiments,the one or more portions of chromosome six is the region spanningpositions 0.92 Mbp and 180 Mbp on chromosome six. In some embodiments,the one or more portions of chromosome six is the region spanningpositions 0.92 Mbp and 22 Mbp on chromosome six. In some embodiments,the positions on chromosome six are in relation to the correspondingpositions on chromosome six in a reference genome. In some embodiments,the reference genome is the Zunla reference genome (Qin, C et al. ProcNatl Acad Sci USA. 2014 Apr. 8; 111(14):5135-40). In some embodiments,the one or more portions of chromosome six is the CCS locus. In someembodiments, the F₁hybrid progeny plants comprising one or more tissuescomprising only a yellow allele at the CCS locus are selected.

Methods of Reducing Propagation of Off-Type Pepper Plants

Certain aspects of the present disclosure relate to methods ofvegetatively propagating any of the chimeric pepper plants describedherein to produce progeny pepper plants that produce pepper fruit havinga yellow and red striped, patched, and/or variegated color. In someembodiments, the method increases the likelihood of producing progenypepper plants that produce pepper fruit having a yellow and red striped,patched, and/or variegated color. In some embodiments, the methodreduces the likelihood of producing off-type progeny pepper plants thatdo not produce pepper fruit having a yellow and red striped, patched,and/or variegated color.

In some embodiments, the method comprises selecting plant material fromthe chimeric pepper plant, and vegetatively propagating the plantmaterial. In some embodiments, the selected plant material is mainbranch material, sympodial shoot material, or material from a branchthat already produces pepper fruit having yellow and red striped,patched, and/or variegated color. In some embodiments, the plantmaterial is main branch material. In some embodiments the plant materialis sympodial shoot material. In some embodiments, the plant material isfrom a branch that already produces pepper fruit having yellow and redstriped, patched and/or variegated color. In some embodiments, the plantmaterial is not axillary shoot material. In some embodiments, the plantmaterial is not from a branch that produces pepper fruit having a singlecolor.

In some embodiments, the method comprises screening plant material fromthe chimeric pepper plant, selecting plant material based upon theresults of the screening, and vegetatively propagating the selectedplant material. In some embodiments, screening plant material comprisesperforming marker analysis on the plant material. In some embodiments,the plant material is one or more of filament, placenta, sepal, petal,ovary, anther, vascular, and stem tissue. In some embodiments, the plantmaterial is anther tissue. In some embodiments, the marker analysis issingle nucleotide polymorphism (SNP) analysis. In some embodiments, themarker analysis indicates the zygosity of one or more chromosomes in theplant material. In some embodiments, the marker analysis indicates thezygosity of chromosome six in the plant material. Examples of markers onchromosome six may include, but are not limited to, CONTIG14444:607,CONTIG3495:1013, CONTIG13973:146, CONTIG15080:278, CAM0000501,CONTIG200085:278, CONTIG1349:1399, CONTIG5713:302, CONTIG1355:1037,CONTIG11146:187, CONTIGCAM0002444, CONTIG8615:1817, CONTIG22248:401,CONTIG3916:1756, CONTIG7457:106, CAM0003794, CONTIG3740:1525,CONTIG462:888, CONTIG2594:540, CAM000413, and the markers described inLefebvre, V. et al. Plant Mol Biol 1998 36(5) 785-789. In someembodiments, the marker is CAM000413. In some embodiments, the markeranalysis indicates the zygosity of one or more portions of chromosomesix in the plant material. In some embodiments, the plant material isselected based upon the plant material being heterozygous for one ormore portions of chromosome six. In some embodiments, the plant materialis selected based upon the plant material being heterozygous for theregion spanning positions 0.92 Mbp and 180 Mbp on chromosome six. Insome embodiments, the plant material is selected based upon the plantmaterial being heterozygous for the region spanning positions 0.92 Mbpand 22 Mbp on chromosome six. In some embodiments, the positions onchromosome six are in relation to the corresponding positions onchromosome six in a reference genome. In some embodiments, the referencegenome is the Zunla reference genome (Qin, C et al. Proc Natl Acad SciUSA. 2014 Apr. 8; 111(14):5135-40). In some embodiments, the plantmaterial is selected based upon the plant material being heterozygous atthe CCS locus.

In some embodiments, the method increases the likelihood of producingprogeny pepper plants that produce pepper fruit having a yellow and redstriped, patched, and/or variegated color. In some embodiments, themethod produces 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% progeny pepperplants that produce pepper fruit having a yellow and red striped,patched, and/or variegated color. In some embodiments, the methodproduces 80% or more progeny pepper plants that produce pepper fruithaving a yellow and red striped, patched, and/or variegated color. Insome embodiments, the method produces 85% or more progeny pepper plantsthat produce pepper fruit having a yellow and red striped, patched,and/or variegated color. In some embodiments, the method produces 90% ormore progeny pepper plants that produce pepper fruit having a yellow andred striped, patched, and/or variegated color. In some embodiments, themethod produces 95% or more progeny pepper plants that produce pepperfruit having a yellow and red striped, patched, and/or variegated color.

In some embodiments, the method reduces the likelihood of producingoff-type progeny pepper plants that do not produce pepper fruit having ayellow and red striped, patched, and/or variegated color. In someembodiments, the method reduces the likelihood of producing off-typeprogeny pepper plants that produce pepper fruit having only a singlecolor. In some embodiments, the off-type progeny pepper plants producepepper fruit having a uniformly yellow color. In some embodiments, themethod produces 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%,9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0% off-type progeny pepperplants. In some embodiments, the method produces 20% or fewer off-typeprogeny pepper plants. In some embodiments, the method produces 15% orfewer off-type progeny pepper plants. In some embodiments, the methodproduces 10% or fewer off-type progeny pepper plants. In someembodiments, the method produces 5% or fewer off-type progeny pepperplants.

Healey Mutant—RY5

‘RY5’ was discovered as a spontaneous mutant in a screening trial ofmutants of pepper variety ‘Healey’ (not patented), conducted in Ontario,Canada in 2016 and in Michigan, United States in 2017. The mutant ‘RY5’was selected based on its vertical red and yellow stripes color andpropagated vegetatively (i.e., asexually).

The pepper plant variety ‘RY5’ has been asexually reproduced by cuttingsat a farm in Ontario, Canada. Pepper plant variety ‘RY5’ isdistinguished from all existing pepper plant varieties by the red andyellow striped fruit of the variety. The distinctive characteristics ofpepper plant variety ‘RY5’ have been found to be stable and aretransmitted to new pepper plants when asexually propagated.

The following botanical description is from fruit grown from plantsvegetatively propagated from the original mutant ‘RY5’ pepper plant.Color references are primarily to the RHS Colour Chart of The RoyalHorticultural Society of London (RHS) (2007 edition). Descriptiveterminology follows the Plant Identification Terminology, An IllustratedGlossary, 2^(nd) edition by James G. Harris and Melinda Woolf Harris,unless where otherwise defined.

Hybrid pepper plant variety ‘RY5’ has the following morphologic andother characteristics:

General:

-   -   Usage: Fresh market    -   Type of culture: Covered cultivation (e.g., in greenhouse)

Plant:

-   -   Seedling (anthocyanin coloration of hypocotyl): Present    -   Stem length: Medium    -   Shortened internode: Absent    -   Internode length: Medium to long    -   Anthocyanin coloration of nodes: Present    -   Intensity of anthocyanin coloration of nodes on stem: Medium to        strong    -   Hariness of nodes on stem: Medium    -   Plant height: Medium to tall    -   Leaf shape: Ovate    -   Undulation of leaf margin: Weak    -   Leaf blistering: Medium to strong    -   Leaf profile in cross section: Moderately convex    -   Leaf glossiness: Medium to strong    -   Peduncle attitude: Semi-dropping    -   Leaf length of blade: Medium to long    -   Leaf width of blade: Medium to broad    -   Intensity of green color of leaf blade: Medium to dark    -   Anthocyanin coloration in anther of flower: Present    -   Time of beginning of flowering: Medium    -   Time of maturity: Medium

Fruit: Images of the ‘RY5’ pepper fruit are presented in FIGS. 7A-7D.

-   -   Color before maturity: Green    -   Intensity of color before maturity: Medium    -   Anthocyanin coloration before maturity: Absent    -   Attidue: Drooping    -   Shape in longitudinal section: Square    -   Shape in cross section (at level of placenta): Angular to        circular    -   Sinuation of pericarp at basal part: Absent or very weak    -   Sinuation of pericarp excluding basal part: Absent or very weak    -   Color at maturity: Red and yellow striped    -   Number of locules: Equally three and four    -   Capsaicin in placenta: Absent    -   Fruit length: Short    -   Fruit width: Broad    -   Ration of length to diameter: Medium    -   Texture of surface of fruit: smooth or very slightly wrinkled    -   Glossiness of fruit: Medium    -   Stalk cavity: Present    -   Depth of stalk cavity: Medium    -   Stalk length: Medium to long    -   Stalk thickness: Medium    -   Calyx aspect: Non-enveloping    -   Shape of apex of fruit: Moderately depressed    -   Depth of interloculary grooves: Medium    -   Thickness of fruit flesh: Medium to thick

Disease/Pest Resistance:

-   -   Tobamovirus pathotype P₉: Resistant    -   Tobamovirus pathotype P₁₋₂: Resistant    -   Tobamovirus pathotype P₁₋₂₋₃: Susceptible    -   Tobamovirus pathotype P₁: Resistant    -   Potato Virus Y pathotype P₀: Susceptible

Comparisons to Most Similar Variety

Table A below compares the characteristic of the mutant pepper plantvariety ‘RY5’ with the most similar variety, ‘Healey’. Column 1 liststhe characteristic, column 2 shows the characteristics for mutant pepperplant variety ‘RY5’, and column 3 shows the characteristics for mostsimilar pepper variety ‘Healey’.

TABLE A Characteristic ‘RY5’ ‘Healey’ Mature fruit color Red and yellowstriped Red

Deposit Information Seed of Pepper Variety ‘RY1’

A deposit of the pepper variety ‘RY1’ is maintained by Enza Zaden BeheerB. V., having an address at Haling 1e, 1602 DB Enkhuizen, Netherlands.Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 2,500 seeds of thesame variety with the National Collection of Industrial, Food and MarineBacteria Ltd. (NCIMB Ltd), Ferguson Building, Craibstone Estate,Bucksburn, Aberdeen, AB21 9YA, United Kingdom.

At least 2500 seeds of pepper variety ‘RY1’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X1. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Tissue Culture of Pepper Variety ‘RY1’

A deposit of tissue culture of the pepper variety ‘RY1’ is maintained byEnza Zaden Beheer B. V., having an address at Haling 1e, 1602 DBEnkhuizen, Netherlands. Access to this deposit will be available duringthe pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to the tissueculture deposit of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

Tissue culture of pepper variety ‘RY1’ were deposited on DATE accordingto the Budapest Treaty in the National Collection of Industrial, Foodand Marine Bacteria Ltd. (NCIMB Ltd), Ferguson Building, CraibstoneEstate, Bucksburn, Aberdeen, AB21 9YA, United Kingdom. The deposit hasbeen assigned NCIMB number Y1. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Seed of Pepper Variety ‘RY2’

A deposit of the pepper variety ‘RY2’ is maintained by Enza Zaden BeheerB. V., having an address at Haling 1e, 1602 DB Enkhuizen, Netherlands.Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 2,500 seeds of thesame variety with the National Collection of Industrial, Food and MarineBacteria Ltd. (NCIMB Ltd), Ferguson Building, Craibstone Estate,Bucksburn, Aberdeen, AB21 9YA, United Kingdom.

At least 2500 seeds of pepper variety ‘RY2’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X2. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Tissue Culture of Pepper Variety ‘RY2’

A deposit of tissue culture of the pepper variety ‘RY2’ is maintained byEnza Zaden Beheer B. V., having an address at Haling 1e, 1602 DBEnkhuizen, Netherlands. Access to this deposit will be available duringthe pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to the tissueculture deposit of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

Tissue culture of pepper variety ‘RY2’ were deposited on DATE accordingto the Budapest Treaty in the National Collection of Industrial, Foodand Marine Bacteria Ltd. (NCIMB Ltd), Ferguson Building, CraibstoneEstate, Bucksburn, Aberdeen, AB21 9YA, United Kingdom. The deposit hasbeen assigned NCIMB number Y2. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Seed of Pepper Variety ‘RY3’

A deposit of the pepper variety ‘RY3’ is maintained by Enza Zaden BeheerB. V., having an address at Haling 1e, 1602 DB Enkhuizen, Netherlands.Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 2,500 seeds of thesame variety with the National Collection of Industrial, Food and MarineBacteria Ltd. (NCIMB Ltd), Ferguson Building, Craibstone Estate,Bucksburn, Aberdeen, AB21 9YA, United Kingdom.

At least 2500 seeds of pepper variety ‘RY3’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X3. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Tissue Culture of Pepper Variety ‘RY3’

A deposit of tissue culture of the pepper variety ‘RY3’ is maintained byEnza Zaden Beheer B. V., having an address at Haling 1e, 1602 DBEnkhuizen, Netherlands. Access to this deposit will be available duringthe pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to the tissueculture deposit of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

Tissue culture of pepper variety ‘RY3’ were deposited on DATE accordingto the Budapest Treaty in the National Collection of Industrial, Foodand Marine Bacteria Ltd. (NCIMB Ltd), Ferguson Building, CraibstoneEstate, Bucksburn, Aberdeen, AB21 9YA, United Kingdom. The deposit hasbeen assigned NCIMB number Y3. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Seed of Pepper Variety ‘RY4’

A deposit of the pepper variety ‘RY4’ is maintained by Enza Zaden BeheerB. V., having an address at Haling 1e, 1602 DB Enkhuizen, Netherlands.Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 2,500 seeds of thesame variety with the National Collection of Industrial, Food and MarineBacteria Ltd. (NCIMB Ltd), Ferguson Building, Craibstone Estate,Bucksburn, Aberdeen, AB21 9YA, United Kingdom.

At least 2500 seeds of pepper variety ‘RY4’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X4. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Tissue Culture of Pepper Variety ‘RY4’

A deposit of tissue culture of the pepper variety ‘RY4’ is maintained byEnza Zaden Beheer B. V., having an address at Haling 1e, 1602 DBEnkhuizen, Netherlands. Access to this deposit will be available duringthe pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to the tissueculture deposit of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

Tissue culture of pepper variety ‘RY4’ were deposited on DATE accordingto the Budapest Treaty in the National Collection of Industrial, Foodand Marine Bacteria Ltd. (NCIMB Ltd), Ferguson Building, CraibstoneEstate, Bucksburn, Aberdeen, AB21 9YA, United Kingdom. The deposit hasbeen assigned NCIMB number Y4. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Seed of Pepper Variety ‘RY5’

A deposit of the pepper variety ‘RY5’ is maintained by Enza Zaden BeheerB. V., having an address at Haling 1e, 1602 DB Enkhuizen, Netherlands.Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 2,500 seeds of thesame variety with the National Collection of Industrial, Food and MarineBacteria Ltd. (NCIMB Ltd), Ferguson Building, Craibstone Estate,Bucksburn, Aberdeen, AB21 9YA, United Kingdom.

At least 2500 seeds of pepper variety ‘RY5’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X5. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Tissue Culture of Pepper Variety ‘RY5’

A deposit of tissue culture of the pepper variety ‘RY5’ is maintained byEnza Zaden Beheer B. V., having an address at Haling 1e, 1602 DBEnkhuizen, Netherlands. Access to this deposit will be available duringthe pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to the tissueculture deposit of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

Tissue culture of pepper variety ‘RY5’ were deposited on DATE accordingto the Budapest Treaty in the National Collection of Industrial, Foodand Marine Bacteria Ltd. (NCIMB Ltd), Ferguson Building, CraibstoneEstate, Bucksburn, Aberdeen, AB21 9YA, United Kingdom. The deposit hasbeen assigned NCIMB number Y5. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Seed of Pepper Variety ‘Maduro’

A deposit of the pepper variety ‘Maduro’ is maintained by Enza ZadenBeheer B. V., having an address at Haling 1e, 1602 DB Enkhuizen,Netherlands. Access to this deposit will be available during thependency of this application to persons determined by the Commissionerof Patents and Trademarks to be entitled thereto under 37 C.F.R. § 1.14and 35 U.S.C. § 122. Upon allowance of any claims in this application,all restrictions on the availability to the public of the variety willbe irrevocably removed by affording access to a deposit of at least2,500 seeds of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

At least 2500 seeds of pepper variety ‘Maduro’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X6. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Seed of Pepper Variety ‘Marinello’

A deposit of the pepper variety ‘Marinello’ is maintained by Enza ZadenBeheer B. V., having an address at Haling 1e, 1602 DB Enkhuizen,Netherlands. Access to this deposit will be available during thependency of this application to persons determined by the Commissionerof Patents and Trademarks to be entitled thereto under 37 C.F.R. § 1.14and 35 U.S.C. § 122. Upon allowance of any claims in this application,all restrictions on the availability to the public of the variety willbe irrevocably removed by affording access to a deposit of at least2,500 seeds of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

At least 2500 seeds of pepper variety ‘Maranello’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X7. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

Seed of Pepper Variety ‘Healey’

A deposit of the pepper variety ‘Healey’ is maintained by Enza ZadenBeheer B. V., having an address at Haling 1e, 1602 DB Enkhuizen,Netherlands. Access to this deposit will be available during thependency of this application to persons determined by the Commissionerof Patents and Trademarks to be entitled thereto under 37 C.F.R. § 1.14and 35 U.S.C. § 122. Upon allowance of any claims in this application,all restrictions on the availability to the public of the variety willbe irrevocably removed by affording access to a deposit of at least2,500 seeds of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

At least 2500 seeds of pepper variety ‘Healey’ were deposited on DATEaccording to the Budapest Treaty in the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom. The deposit has been assigned NCIMB number X8. Access to thisdeposit will be available during the pendency of this application topersons determined by the Commissioner of Patents and Trademarks to beentitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Uponallowance of any claims in this application, all restrictions on theavailability to the public of the variety will be irrevocably removed.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

The present disclosure will be more fully understood by reference to thefollowing Examples. They should not, however, be construed as limitingthe scope of the present disclosure.

EXAMPLES Example 1 Identification and Characterization of RY1 PepperPlant

Pepper is an important and valuable field crop, and there is a continuedneed for developing new and interesting hybrid peppers. To this end, aninbred pepper plant that produces yellow fruit (pepper variety‘OP.1745’) and an inbred pepper plant that produces red fruit (peppervariety ‘OP.2023’) were crossed, and the resulting seeds were planted tocreate a population of F₁ hybrid plants. The resulting F₁ hybrid plants,commercially available as pepper variety ‘Maduro’, were grown, and aselection process was initiated to identify mutant plants that producedfruit with alternating yellow and red stripes or patches (‘Maduro’produces uniform, single-colored red fruit when mature).

One of these mutant plants (RY1) was selected for furthercharacterization. This plant produced fruit with alternating, irregularyellow and red stripes/patches, with each fruit having a unique colorpattern (FIG. 1). The irregular fruit color distribution pattern wasalso observed across the fruit wall from outside to inside, and theoutside and inside of the fruit could have the same or differentcoloring (FIG. 2). Aside from the unique coloring of the fruit, RY1 wasphenotypically identical to ‘Maduro’. Interestingly, while selfingprogeny of ‘Maduro’ segregated for fruit color in a typical Mendelian3:1 (red to yellow) ratio, selfing progeny of RY1 did not segregate in atypical Mendelian ratio, only bearing off-type, uniformly yellow fruit.

To further characterize the RY1 plants, DNA marker profiles weregenerated for the inbred parent plants ‘OP.1745’ and ‘OP.2023’,‘Maduro’, and RY1, using markers distributed over all 12 chromosomes ofthe plants. To gain a clearer understanding of the genetic profile ofRY1, marker profiles were generated for multiple leaf tissues, as wellas multiple portions of both the yellow and red parts of the fruit.Interestingly, the marker profile for some of the portions of RY1 (e.g.,the portions taken from the red part of the fruit) were identical to‘Maduro’, while the marker profile for other portions of RY1 (e.g., theportions taken from the yellow part of the fruit, and some, but not all,of the leaf samples) showed an aberrant profile with respect to themarkers located on the upper portion of chromosome six, between basepair positions 927,315 and 22,373,591 (FIG. 3). At these positions, themarker profiles were similar to the marker profiles for ‘OP.1745’ (theinbred parent pepper plant that produces yellow fruit), and not‘Maduro’. Markers at positions 180,205,121 and 190,879,102 on chromosomesix were identical to ‘Maduro’ for all tissues tested. Without wishingto be bound by theory, these data suggested that some of the tissues ofRY1 had lost the upper portion of chromosome six that was derived from‘OP.2023’ (the inbred parent pepper plant that produces red fruit),indicating that these tissues may have been hemizygous for the upperportion of chromosome six.

The Capsanthin-Capsorubin Synthase (CCS) gene provides peppers withtheir color (See e.g., Thorup, T. A. et al. Proc. Natl. Acad. Sci. USA.2000 Oct. 10; 97(21):11192-7). Red or yellow fruit color is determinedby the presence or absence of a functional CCS gene, respectively. TheCCS gene is located on the upper portion of chromosome six, between basepair positions 927,315 and 22,373,591. The yellow allele at the CCSlocus harbors C to A and T to C polymorphisms in the nucleic acidencoding the 3′ UTR of the CCS transcript, and a marker, CAM000413, isable to detect these polymorphisms (FIG. 4). The CAM000413 marker wasused to identify the alleles of the CCS locus in 9 samples (3independent samples of each organ/tissue from 3 plants) of variousorgan/tissue types for both ‘Maduro’ and RY1 plants. Table 1 describesthe results of this analysis.

TABLE 1 CAM000413 marker analysis for tissues from ‘Maduro’ and RY1‘Maduro’ RY1 Yellow Red and Red Yellow Red and Red allele yellow alleleallele yellow allele Tissue only alleles only only alleles only Seed 0 90 9 0 0 Epidermal 0 9 0 9 0 0 peel fruit Filament 0 9 0 7 2 0 Pericarp 09 0 6 (yellow 3 (red 0 part) part) Septum 0 9 0 9 0 0 Placenta 0 9 0 1 80 Sepal 0 9 0 6 3 0 Petal 0 9 0 8 1 0 Ovary 0 9 0 4 5 0 Anther 0 9 0 1 80 Leaf - 0 9 0 8 1 0 middle part Leaf - 0 9 0 9 0 0 side part vascular 09 0 5 3 0 Stem 0 9 0 5 4 0

As expected, all 9 samples of each of the organ/tissue types tested for‘Maduro’ contained both a red and yellow allele at the CCS locus. Incontrast to ‘Maduro’, some of the organ/tissue types of RY1 containedboth a red and yellow allele at the CCS locus (e.g., the anther andplacenta tissues), while other tissue/organ types appeared to onlycontain a yellow allele at the CCS locus (e.g., the epidermal peel,filament, septum, petal, and leaf parts tissues). Additionally, someorgan/tissue types tested were a more even distribution of containingboth a red and yellow allele at the CCS locus, and only a yellow alleleat this locus (e.g., the pericarp, sepal, ovary, vascular and stemtissues). Interestingly, the 9 seed samples taken from RY1 onlycontained the yellow allele at the CCS locus. Taken together, theseresults revealed that the RY1 plant was a chimeric pepper plant havingsome tissues containing both the yellow and red allele at the CCS locus,and other tissues with only the yellow allele at the CCS locus.

Example 2 Identification and Characterization of RY2 Pepper Plant

Field trials were initiated to identify other inbred pepper plantcrosses that would give rise to mutant F₁ hybrid pepper plants thatproduce yellow and red striped, patched or variegated pepper fruit. Assuch, an inbred pepper plant that produces yellow fruit (pepper variety‘OP.1745’) and an inbred pepper plant that produces red fruit (peppervariety ‘OP.1755’) were crossed, and the resulting seeds were planted tocreate a population of F₁ hybrid plants. The resulting F₁ hybrid plants,commercially available as pepper variety ‘Marinello’, were grown, and aselection process was initiated to identify mutant plants of this F₁hybrid that produced fruit with alternating yellow and red stripes orpatches (‘Marinello’ produces uniform, single-colored red fruit whenmature). Another mutant F₁ hybrid plant (RY2) was found to produceyellow and red striped/patched pepper fruit. This plant produced fruitwith alternating, irregular yellow and red stripes/patches (FIG. 5),similar to the fruit produced by RY1. Similar to the experimentsdescribed above, the CAM000413 marker was used to identify the allelesof the CCS locus in four samples of red and yellow pericarp tissueobtained from the fruit of the RY2 plants. Table 2 describes the resultsof this analysis.

TABLE 2 CAM000413 marker analysis for tissues from RY2 RY2 Red andyellow Yellow Red allele Tissue allele only alleles only UndeterminedPericarp 1 (yellow 3 (red part) 0 1 (red part) part) 1 (yellow 2 (yellowpart) part)

Three of the four samples taken from the red portion of the pericarptissue contained the CAM000413 marker for both the red and yellowalleles, and one of the four samples taken from the yellow portion ofthe pericarp tissue contained the CAM000413 marker for only the yellowallele, in agreement with the results observed for sample from RY1(Table 1 above). Unexpectedly, one of the four samples taken from theyellow tissue contained the CAM000413 marker for both the red and yellowalleles, and no reliable results could be obtained for the other yellowpericarp samples. This suggests that there may have been crosscontamination from the red portion of the pericarp tissue in the yellowsamples, or the DNA quality of the samples was low.

A more extensive analysis of the CAM000413 marker was used to identifythe alleles of the CCS locus in 9 samples (3 independent samples of eachorgan/tissue from 3 plants) of various organ/tissue types for both‘Maranello’ and RY2 plants to better understand these plants. Table 3describes the results of this analysis.

TABLE 3 CAM000413 marker analysis for tissues from ‘Maranello’ and RY2‘Maranello’ RY2 Yellow Red and Red Yellow Red and Red allele yellowallele allele yellow allele Tissue only alleles only only alleles onlySeed 0 9 0 1 8 0 Epidermal 0 9 0 0 9 0 peel fruit Filament 0 9 0 0 9 0Pericarp 0 9 0 5 (Yellow 4 (Red 0 part) part) Septum 0 9 0 0 9 0Placenta 0 9 0 0 9 0 Sepal 0 9 0 0 9 0 Petal 0 9 0 0 9 0 Ovary 0 9 0 0 90 Anther 0 9 0 0 9 0 Leaf - 0 9 0 0 9 0 middle part Leaf - 0 9 0 1 8 0side part vascular 0 9 0 0 9 0 Stem 0 9 0 0 9 0

Similar to what was observed for ‘Maduro’ and RY1, all tissue samples of‘Maranello’ contain both the red and the yellow allele at the CCs locus,whereas in RY2 some samples show only the yellow allele and others boththe yellow and the red alleles in different tissues.

Example 3 Identification and Characterization of RY3 Pepper Plant

A third mutant pepper plant was identified that produced yellow and redstriped/patched/variegated pepper fruit (RY3). This pepper plant was amutant of an F₁ hybrid pepper plant, commercially available as peppervariety ‘Waltz’. Table 4 shows marker analysis performed on the F₁hybridplants ‘Waltz’, ‘Maduro’, and ‘Maranello’, as well as the parentalplants of ‘Maduro’ and ‘Maranello’ (‘OP.1745’, ‘OP.2023’, and‘OP.1755’).

TABLE 4A Marker analysis of F₁ hybrid and parental pepper plants Marker(CAM0000) Plant: 402 408 413 442 448 450 459 468 469 471 472 473 491‘OP.2023’ 0202 0202 0101 0202 0101 0101 0202 0202 0202 0202 0202 01010202 ‘OP.1745’ 0101 0202 0202 0101 0202 0202 0202 0101 0202 0101 02020202 0505 ‘Maduro’ 0102 0202 0102 0102 0102 0102 0202 0102 0202 01020202 0102 0205 ‘OP.1755’ 0202 0202 0101 0101 0101 0101 0202 0202 02020202 0202 0101 0202 ‘OP.1745’ 0101 0202 0202 0101 0202 0202 0202 01010202 0101 0202 0202 0505 ‘Maranello’ 0102 0202 0102 0101 0102 0102 02020102 0202 0102 0202 0102 0205 ‘Waltz’ 0202 0202 0102 0202 0101 0102 02020102 0202 0202 0202 0202 9999 0101 = homozygous for allele 1 0202 =homozygous for allele 2 0102 = heterozygous for allele 1 and 2 0505 =homozygous for allele 5 9999 = unknown marker score

As indicated by the marker analysis in Table 4, the F₁ hybrid pepperplant ‘Waltz’ did not share common parent plants with either ‘Maduro’ or‘Maranello’, suggesting that the mutant pepper plant derived from‘Waltz’ was yet another distinctive pepper plant that produced yellowand red striped fruit.

An analysis of the CAM000413 marker was used to identify the alleles ofthe CCS locus in 9 samples (3 independent samples of each organ/tissuefrom 3 plants) of various organ/tissue types for both ‘Waltz’ and RY3plants. Table 4B describes the results of this analysis.

TABLE 4B CAM000413 marker analysis for tissues from ‘Waltz’ and RY3‘Waltz’ RY3 Yellow Red and Red Yellow Red and Red allele yellow alleleallele yellow allele Tissue only alleles only only alleles only Seed 0 90 1 8 0 Epidermal 0 9 0 3 6 0 peel fruit Filament 0 9 0 4 5 0 Pericarp 09 0 4 (yellow 5 (red 0 part) part) Septum 0 9 0 6 3 0 Placenta 0 9 0 0 90 Sepal 0 9 0 8 1 0 Petal 0 9 0 4 5 0 Ovary 0 9 0 0 9 0 Anther 0 9 0 9 00 Leaf - 0 9 0 8 1 0 middle part Leaf - 0 9 0 7 2 0 side part vascular 09 0 2 6 0 Stem 0 9 0 0 9 0

Similar to what was observed for ‘Maduro’ and RY1, all tissue samples of‘Wlatz’ contain both the red and the yellow allele at the CCs locus,whereas in RY3 some samples show only the yellow allele and others boththe yellow and the red alleles in different tissues.

Example 4 Identification and Characterization of RY4 Pepper Plant

A fourth mutant pepper plant was identified that produced yellow and redstriped/patched/variegated pepper fruit (RY4). This pepper was an F₃hybrid pepper derived from a breeding scheme using a proprietaryparental pepper line that produced red fruit (‘Rubinex’ x ‘OP.1091’ xOP.1747’) crossed with a proprietary parental pepper line that producedyellow fruit (‘OP.1033 x (Br CTx378) x ‘Ramiro). F₃ hybrid plants wereproduced from this breeding scheme, the resulting F₃ hybrid plants weregrown, and a selection process was initiated to identify mutant plantsthat produced fruit with alternating yellow and red stripes or patches.One of these mutant plants (RY4) produced fruit with alternating,irregular yellow and red stripes/patches (FIG. 6).

Example 5 Identification and Characterization of RY5 Pepper Plant

A fifth mutant pepper plant was identified that produced yellow and redstriped/patched/variegated pepper fruit (RY5). A pepper plant varietythat produces yellow fruit (pepper variety ‘OP.1745’) and a pepper plantvariety that produces red fruit (pepper variety ‘OP.0900’) were crossed,and the resulting seeds were planted to create a population of F₁ hybridplants. The resulting F₁ hybrid plants, commercially available as peppervariety ‘Healey’, were grown, and a selection process was initiated toidentify mutant plants that produced fruit with alternating yellow andred stripes or patches. One of these mutant plants (RY5) produced fruitwith alternating, irregular yellow and red stripes/patches (FIG. 7A-D).Table 5 shows marker analysis performed on the F₁hybrid plants ‘Healey’,as well as the parental plants of ‘Healy’ (‘OP.1745’ and ‘OP.0900’).

TABLE 5 Marker analysis of F₁ hybrid and parental pepper plants Marker(CAM0000) Plant: 402 408 413 442 448 450 459 468 469 471 472 473 491‘OP.0900 0202 0202 0101 0101 0101 0101 0202 0202 0202 0202 0202 01010202 ‘OP.1745’ 0101 0202 0202 0101 0202 0202 0202 0101 0202 0101 02020202 0505 ‘Healey’ 0102 0202 0102 0101 0102 0102 0202 0102 0202 01020202 0102 0205 0101 = homozygous for allele 1 0202 = homozygous forallele 2 0102 = heterozygous for allele 1 and 2 0505 = homozygous forallele 5 9999 = unknown marker score

An analysis of the CAM000413 marker was used to identify the alleles ofthe CCS locus in 9 samples (3 independent samples of each organ/tissuefrom 3 plants) of various organ/tissue types for both ‘Healey’ and RY5plants. Table 6 describes the results of this analysis.

TABLE 6 CAM000413 marker analysis for tissues from ‘Healey’ and RY5‘Healey’ RY5 Yellow Red and Red Yellow Red and Red allele yellow alleleallele yellow allele Tissue only alleles only only alleles only Seed 0 90 6 3 0 Epidermal 0 9 0 5 4 0 peel fruit Filament 0 9 0 0 9 0 Pericarp 09 0 4 (yellow 5 (red 0 part) part) Septum 0 9 0 7 2 0 Placenta 0 9 0 0 90 Sepal 0 9 0 8 1 0 Petal 0 9 0 3 6 0 Ovary 0 9 0 1 8 0 Anther 0 9 0 7 20 Leaf - 0 9 0 8 1 0 middle part Leaf - 0 9 0 9 0 0 side part vascular 09 0 1 8 0 Stem 0 9 0 0 9 0

Similar to what was observed for ‘Maduro’ and RY1, all tissue samples of‘Healey’ contain both the red and the yellow allele at the CCs locus,whereas in RY5 some samples show only the yellow allele and others boththe yellow and the red alleles in different tissues.

Example 6 Vegetative Propagation of RY1 Chimeric Pepper Plant

The yellow and red striped pepper phenotype was not found in the progenyof the RY1 chimeric pepper plant. As indicated in Table 1, the seedsproduced from these plants only contained the yellow allele at the CCSlocus, and peppers produced by plants grown from these seeds always boreoff-type, uniformly yellow fruit. The yellow and red striped pepperphenotype was found, nonetheless, in plants produced via cuttings fromthe RY1 chimeric pepper plant. However, the yellow and red pepperphenotype was somewhat unstable, as some main or side branches wouldspontaneously bear only off-type, uniformly yellow fruit. Once a branchbore one off-type, uniformly yellow fruit, all subsequent fruit born onthat branch were off-type, uniformly yellow. The frequency with whichcuttings resulted in plants with off-type producing branches ranged from5-35%. As such, experiments were conducted to determine whetherpropagating particular plant material via cuttings could reduce ormitigate the number/frequency of plants with branches bearing off-type,yellow fruit.

To this end, cuttings were taken from the main branch, as well aslateral (sympodial branching) and axillary shoots, and the cuttings werevegetatively propagated. Next, the number of plants producing off-type,uniformly yellow fruit was assessed. Vegetatively propagated plantsderived from axillary shoots produced a high frequency (25-35%) ofplants producing off-type fruit, while surprisingly, vegetativelypropagated plants derived from lateral shoots had a much lower frequency(5-15%) of plants producing off-type, uniformly yellow fruit.Additionally, vegetatively propagated plants from main branch materialshowed a low frequency of plants producing off-type fruit. These resultssuggested that vegetatively propagating plants by cuttings from the mainbranch or lateral shoots could reduce the frequency of off-type plants,with lateral shoots providing an unexpected 1.7 to 7 fold reduction inoff-type plants when compared to axillary shoots.

Example 7 Vegetatively Propagating Plant Material Based on MarkerAnalysis

Another method of reducing the number of plants producing off-type,uniformly yellow fruit during vegetative propagation is to performmarker analysis on the plant material. Cuttings are taken from achimeric plant producing yellow and red striped pepper fruit, and theplant material is subjected to marker analysis. Any marker capable ofdetecting a genetic difference (e.g., a single nucleotide polymorphism(SNP) at one or more chromosomal locations) between the parental plantslocated on chromosome six is used (e.g., CCS locus-linked markers).Plant material that is heterozygous for the tested positions (i.e.,carries chromosome six from both parental plants) is selected, as thismaterial gives the highest likelihood of propagating plants that produceyellow and red striped/patched pepper fruit. Plant material that is notheterozygous at the tested positions is discarded, as this materialgives rise to plants that produce off-type, uniformly yellow fruit.

While any one or more plant materials may be tested and propagated whengiving a heterozygous marker profile for chromosome six as describedabove, anthers provide a robust and reliable source of plant materialthat diminishes the frequency of producing off-type plants. Anthers aretested at the stage when the first flowers appear on the branches.Marker analysis is performed on the anther tissue, and anthers that arenot heterozygous for the tested positions on chromosome six indicate abranch that will not produce yellow and red striped fruit. Cuttings fromthese branches may be discarded. Anthers that are heterozygous for thetested positions on chromosome six indicate a branch that will produceyellow and red striped fruit, and Cuttings from these branches may beused for vegetative propagation. Using this protocol can reduce thenumber/frequency of plants producing off-type fruit, producing moretrue-to-type plants.

Example 8 Propagating Chimeric Pepper Plants Via the BBM Method

An additional method to propagate pepper plants producing pepper fruithaving a red and yellow striped color is to regenerate plants fromsomatic embryos generated from chimeric pepper plants. To this end, achimeric pepper plant producing pepper fruit having a red and yellowstriped color is transformed with a vector to ectopically express thetranscription factor BABY BOOM AP2/ERF from Brassica napus (See e.g.,Heidmann et al. Plant Cell Rep 30:1107-1115 and/or WO2011003471).Transient activation of BABY BOOM in the progeny plants induces cellregeneration and is used to produce somatic embryos. Pepper plants arethen regenerated from the somatic embryos produced from the transformedchimeric pepper plants (See e.g., Heidmann 2015 PhD thesis WageningenAgricultural University ISBN 978-94-6257-466-3). These pepper plants,regenerated using the BBM transformation protocol, produce pepper fruithaving a yellow and red striped color.

1-36. (canceled)
 37. A chimeric pepper plant comprising a first alleleat a Capsanthin-Capsorubin Synthase (CCS) locus, wherein the firstallele comprises SEQ ID NO: 4, and a second allele at the CCS locus,wherein the second allele comprises SEQ ID NO: 5, wherein the chimericpepper plant produces pepper fruit having yellow and red striped color.38. The chimeric pepper plant of claim 37, wherein the chimeric pepperplant comprises one or more first tissues comprising only the secondallele and one or more second tissues comprising the first allele andthe second allele, wherein the first tissue is one or more tissuesselected from the group consisting of epidermal peel tissue, septumtissue, sepal tissue, anther tissue, and leaf tissue, and wherein thesecond tissue is one or more tissues selected from the group consistingof filament tissue, placenta tissue, petal tissue, ovary tissue,vascular tissue, and stem tissue.
 39. The chimeric pepper plant of claim37, wherein the chimeric pepper plant produces chimeric pericarp havingone or more regions having a yellow color and comprising only the secondallele and one or more regions having a red color and comprising thefirst allele.
 40. A plant part from the chimeric pepper plant of claim37, wherein the plant part is a leaf, a seed, a fruit, a cell, or aportion thereof.
 41. The plant part of claim 40, wherein the plant partis a fruit.
 42. A chimeric pepper plant having all the physiological andmorphological characteristics of the chimeric pepper plant of claim 37.43. A plant part from the chimeric pepper plant of claim 42, wherein theplant part is a leaf, a seed, a fruit, a cell, or a portion thereof. 44.The plant part of claim 43, wherein the plant part is a fruit.
 45. Apollen grain or an ovule of the chimeric pepper plant of claim
 37. 46. Aprotoplast produced from the chimeric pepper plant of claim
 37. 47. Atissue culture produced from protoplasts or cells from the chimericpepper plant of claim 37, wherein the protoplasts or cells are producedfrom a plant part selected from the group consisting of leaf, anther,pistil, stem, petiole, root, root tip, fruit, seed, flower, cotyledon,hypocotyl, embryo, and meristematic cell.
 48. A chimeric pepper plantproduced from a cross of a first pepper plant with a second pepperplant, wherein the first pepper plant comprises a first allele at aCapsanthin-Capsorubin Synthase (CCS) locus, wherein the first allelecomprises SEQ ID NO: 4, and the second pepper plant comprises a secondallele at the CCS locus, wherein the second allele comprises SEQ ID NO:5; wherein the first pepper plant produces pepper fruit having a redcolor and the second pepper plant produces pepper fruit having a yellowcolor; wherein the cross produces the chimeric pepper plant; and whereinthe chimeric pepper plant comprises one or more first tissues comprisingonly the second allele and one or more second tissues comprising thefirst allele and the second allele.
 49. The chimeric pepper plant ofclaim 48, wherein the first tissue is one or more tissues selected fromthe group consisting of epidermal peel tissue, septum tissue, sepaltissue, anther tissue, and leaf tissue.
 50. A seed produced from thechimeric pepper plant of claim 48, wherein the seed comprises the firstallele and the second allele.
 51. A chimeric pepper plant produced bygrowing the seed of claim
 50. 52. The chimeric pepper plant of claim 51,wherein the chimeric pepper plant produces pepper fruit having red andyellow striped color.
 53. A plant part from the chimeric pepper plant ofclaim 48, wherein the plant part is a leaf, a seed, a fruit, a cell, ora portion thereof.
 54. The plant part of claim 53, wherein the plantpart is a fruit.
 55. A pepper plant having all the physiological andmorphological characteristics of the pepper plant of claim
 48. 56. Aplant part from the pepper plant of claim 55, wherein the plant part isa leaf, a seed, a fruit, a cell, or a portion thereof.
 57. The plantpart of claim 56, wherein the plant part is a fruit.
 58. A pollen grainor an ovule of the chimeric pepper plant of claim
 48. 59. A protoplastproduced from the chimeric pepper plant of claim
 48. 60. A tissueculture produced from protoplasts or cells from the chimeric pepperplant of claim 48 wherein the protoplasts or cells are produced from aplant part selected from the group consisting of leaf, anther, pistil,stem, petiole, root, root tip, fruit, seed, flower, cotyledon,hypocotyl, embryo, and meristematic cell.
 61. A method of vegetativelypropagating a chimeric pepper plant that produces pepper fruit havingyellow and red striped color, wherein the chimeric pepper plantcomprises a first allele at a Capsanthin-Capsorubin Synthase (CCS)locus, wherein the first allele comprises SEQ ID NO: 4, and a secondallele at the CCS locus, wherein the second allele comprises SEQ ID NO:5, the method comprising: a) screening plant material from the chimericpepper plant; b) selecting plant material based upon the results of thescreening; and c) vegetatively propagating the selected plant materialto asexually reproduce a chimeric pepper plant that produces pepperfruit having yellow and red striped color.
 62. The method of claim 61,wherein the plant material is selected from the group consisting of asympodial shoot, an axillary shoot, a lateral shoot, a main branch, anda branch that produces pepper fruit having yellow and red striped color.63. The method of claim 62, wherein the plant material is a main branchor a branch that produces pepper fruit having yellow and red stripedcolor.
 64. The method of claim 61, wherein the screening of step (a)comprises a single nucleotide polymorphism (SNP) analysis.
 65. Themethod of claim 64, wherein the SNP analysis is performed on plantmaterial selected from the group consisting of filament material,placenta material, sepal material, petal material, ovary material,anther material, vascular material, and stem material.
 66. The method ofclaim 65, wherein the SNP analysis is performed on an anther material.67. The method of claim 64, wherein the second allele is identified by aC at a position corresponding to nucleotide 49 of SEQ ID NO: 5 and an Aat a position corresponding to nucleotide 85 of SEQ ID NO:
 5. 68. Themethod of claim 61, wherein the selecting of step (b) comprisesselecting plant material based upon the plant material beingheterozygous for the region spanning positions 0.92 Mbp and 180 Mbp onchromosome 6, or wherein the selecting of step (b) comprises selectingplant material based upon the plant material being heterozygous for theregion spanning positions 0.92 Mbp and 22 Mbp on chromosome
 6. 69. Themethod of claim 61, wherein the selecting of step (b) comprisesselecting plant material based on the plant material being heterozygousat the CCS locus.
 70. The method of claim 69, wherein the selected plantmaterial has a first allele at the CCS locus, wherein the first allelecomprises SEQ ID NO: 4, and a second allele at the CCS locus, whereinthe second allele comprises SEQ ID NO:
 5. 71. The method of claim 61,wherein the method reduces the likelihood of producing off-type progenypepper plants that produce pepper fruit having only a single color ascompared to seed based propagation of the chimeric pepper plant.
 72. Themethod of claim 71, wherein the method produces off-type progeny pepperplants that produce pepper fruit having only a single color at afrequency in the range of 35% to 5% or less.
 73. The method of claim 72,wherein the method produces off-type progeny pepper plants that producepepper fruit having only a single color at a frequency in the range of15% to 5% or less.
 74. The method of claim 61, wherein the methodincreases the likelihood of producing progeny pepper plants that producepepper fruit having a yellow and red striped color as compared to seedbased propagation of the chimeric pepper plant.
 75. The method of claim74, wherein the method produces progeny pepper plants that producepepper fruit having a yellow and red striped color at a frequency in therange of 65% to 95% or more.
 76. The method of claim 75, wherein themethod produces progeny pepper plants that produce pepper fruit having ayellow and red striped color at a frequency in the range of 85% to 95%or more.
 77. A pepper plant produced by the method of claim 61.